Use of anaerobic green fluorescent protein versus green fluorescent protein as reporter in lactic acid bacteria
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| 008 | 210128e20150801xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00253-015-6770-3 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6770-3 | ||
| 245 | 0 | 0 | |a Use of anaerobic green fluorescent protein versus green fluorescent protein as reporter in lactic acid bacteria |h [Elektronische Daten] |c [José Landete, Susana Langa, Concepción Revilla, Abelardo Margolles, Margarita Medina, Juan Arqués] |
| 520 | 3 | |a Lactic acid bacteria (LAB) are commonly used in the production of fermented and probiotic foods. Development of molecular tools to discriminate the strains of interest from the endogenous microbiota in complex environments like food or gut is of high interest. Green fluorescent protein (GFP)-like chromophores strictly requires molecular oxygen for maturation of fluorescence, which restrict the study of microorganisms in low-oxygen environments. In this work, we have developed a noninvasive cyan-green fluorescent based reporter system for real-time tracking of LAB that is functional under anoxic conditions. The evoglow-Pp1 was cloned downstream from the promoters d-alanyl-d-alanine carboxypeptidase and elongation factor Tu of Lactobacillus reuteri CECT925 using pNZ8048 and downstream of the lactococcal P1 promoter using pT1NX. The classical gfp was also cloned in pT1NX. These recombinant expression vectors were electroporated into Lactococccus, Lactobacillus, and Enterococcus strains with biotechnological and/or probiotic interests to assess and compare their functionality under different conditions of oxygen and pH. The expression was analyzed by imaging and fluorometric methods as well as by flow cytometry. We demonstrate that reporter systems pNZ:TuR-aFP and pT1-aFP are two versatile molecular markers for monitoring LAB in food and fecal environments without the potential problems caused by oxygen and pH limitations, which could be exploited for in vivo studies. Production of the fluorescent protein did not disturb any important physiological properties of the parental strains, such as growth rate, reuterin, or bacteriocin production. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Fluorescent proteins |2 nationallicence | |
| 690 | 7 | |a Plasmid |2 nationallicence | |
| 690 | 7 | |a Anaerobic conditions |2 nationallicence | |
| 690 | 7 | |a Lactic acid bacteria |2 nationallicence | |
| 700 | 1 | |a Landete |D José |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | |
| 700 | 1 | |a Langa |D Susana |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | |
| 700 | 1 | |a Revilla |D Concepción |u Departamento de Biotecnología, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | |
| 700 | 1 | |a Margolles |D Abelardo |u Instituto de Productos Lácteos de Asturias, CSIC, Paseo Río Linares s/n, 33300, Villaviciosa, Asturias, Spain |4 aut | |
| 700 | 1 | |a Medina |D Margarita |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | |
| 700 | 1 | |a Arqués |D Juan |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/16(2015-08-01), 6865-6877 |x 0175-7598 |q 99:16<6865 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6770-3 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6770-3 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Landete |D José |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Langa |D Susana |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Revilla |D Concepción |u Departamento de Biotecnología, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Margolles |D Abelardo |u Instituto de Productos Lácteos de Asturias, CSIC, Paseo Río Linares s/n, 33300, Villaviciosa, Asturias, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Medina |D Margarita |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Arqués |D Juan |u Departamento de Tecnología de Alimentos, INIA, Carretera de La Coruña Km 7, 28040, Madrid, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/16(2015-08-01), 6865-6877 |x 0175-7598 |q 99:16<6865 |1 2015 |2 99 |o 253 | ||